Nijmegen Breakage Syndrome fibroblasts and iPSCs: cellular models for uncovering disease-associated signaling pathways and establishing a screening platform for anti-oxidants

Abstract

Nijmegen Breakage Syndrome (NBS) is associated with cancer predisposition, premature aging, immune deficiency, microcephaly and is caused by mutations in the gene coding for NIBRIN (NBN) which is involved in DNA damage repair. Dermal-derived fibroblasts from NBS patients were reprogrammed into induced pluripotent stem cells (iPSCs) in order to bypass premature senescence. The influence of antioxidants on intracellular levels of ROS and DNA damage were screened and it was found that EDHB-an activator of the hypoxia pathway, decreased DNA damage in the presence of high oxidative stress. Furthermore, NBS fibroblasts but not NBS-iPSCs were found to be more susceptible to the induction of DNA damage than their healthy counterparts. Global transcriptome analysis comparing NBS to healthy fibroblasts and NBS-iPSCs to embryonic stem cells revealed regulation of P53 in NBS fibroblasts and NBS-iPSCs. Cell cycle related genes were down-regulated in NBS fibroblasts. Furthermore, oxidative phosphorylation was down-regulated and glycolysis up-regulated specifically in NBS-iPSCs compared to embryonic stem cells. Our study demonstrates the utility of NBS-iPSCs as a screening platform for anti-oxidants capable of suppressing DNA damage and a cellular model for studying NBN de-regulation in cancer and microcephaly.

Primary Microcephaly with Novel Variant of MCPH1 Gene in Twins: Both Manifesting in Childhood at the Same Time with Hashimoto's Thyroiditis

“…In the twins, on the basis of clinical and laboratory investigations, the etiologic diagnosis was circumscribed to monogenic cause of microcephaly among the group of autosomal recessive primary microcephaly, and the differential diagnosis was made with other disorders and/or syndromes in which microcephaly is the presenting sign, including Seckel's syndrome (microcephalic dwarfism type 1), 19,20 the α thalassemia mental retardation X-linked syndrome, 21,22 and the Nijmegen breakage syndrome. 23,24 Several genes and protein products have been identified in patients with autosomal recessive primary microcephaly. Eighteen MCPH loci (MCPH1_MCPH18) were mapped and contain the following genes: Microcephalin, WDR62, CDK5RAP2, CASC5, ASPM, CENPJ, STIL, CEP135, ZNF335, PHC1, CDK6,CENPE, SASS6, MFSD2A, ANKLE2, CIT, and WDFY3.…”

Evidence for a pre-malignant cell line in a skin biopsy from a patient with Nijmegen breakage syndrome

“…Moreover, these cells could be cloned. This is in contrast to NBS skin fibroblasts, which grow slowly in culture, enter early into senescence and have an extremely low reprogramming efficiency [ 13 ]. There are strong arguments for the assumption that the derivative chromosome is the crucial, primary event for the selective advantage of this cell line against the NBS background.…”

“…NBS skin fibroblasts grow slowly in culture and enter early into senescence. Consequently, they have a low reprogramming efficiency into induced pluripotent stem cells (iPSCs) [ 13 ]. The iPSCs show, amongst others, numerous chromosomal aberrations, a delayed response to DSB induction, slower growth rate, and a reduced apoptotic response to stress [ 14 ].…”

Bilirubin-Induced Neurological Damage: Current and Emerging iPSC-Derived Brain Organoid Models

“…Further expansions in iPSC research have increasingly revealed the multifaceted use of these cells in modeling various diseases in vitro [ 17 , 18 , 19 , 20 , 100 ]. With the development of iPSCs, researchers have been able to replicate many diseases, including Parkinson’s disease, Nijmegen Breakage Syndrome, and Alzheimer’s disease, all by generating different types of cells that mimic the in vivo environment very closely [ 19 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 ]. For instance, deriving iPSCs from patients with genetic-based neurological conditions and differentiating them into neurons opens up more possibilities to closely observe the pathological mechanisms underlying the disease in vitro [ 106 , 110 , 111 , 112 ].…”